The Protein Man's Blog | A Discussion of Protein Research

The Role of the Toll (like receptor)

Posted by The Protein Man on Jan 19, 2021 4:00:00 PM


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Topics: Protein Estimation

Back to Basics: Western Blot 101

Posted by The Protein Man on Dec 29, 2020 1:00:00 PM

Western blotting or immunoblotting is an indispensable technique, almost every published paper in area of molecular cell biology uses western blotting for detecting specific proteins in a sample of tissue homogenate or cell lysates. Western blotting combines resolving power of polyacrylamide gel electrophoresis (PAGE) or SDS-PAGE and specificity of antibodies to detect target proteins.  Proteins are resolved on the basis of their molecular weight in SDS-PAGE and transferred from the polyacrylamide gel onto the membranes (Nitrocellulose or PVDF), which creates an exact replica of the protein separation pattern on the membrane. After transferring the proteins to the membrane, the membrane needs ‘blocking’ to ‘block’ non-specific binding sites on the surface of the membrane. Blocking is usually performed with Bovine Serum Albumin, Skimmed milk or purified milk Casein.

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Topics: Protein Purification, Western Blotting, Protein Electrophoresis, Protein Estimation, Sample Clean Up, Protease Inhibitors, Protein Extraction, Protein Detection

Guess what??? Immobilized Papain Generates Fab Fragments of Cluster A Monoclonal Antibodies for Validation of Inner Domain of HIV-1 gp120?

Posted by The Protein Man on Aug 19, 2020 3:15:00 PM


The lab of Dr. William D. Tolbert and Dr. Neelakshi Gohain use Papain-linked agarose resin to create combinations of purified Fabs of Cluster A monoclonal antibodies (A32 mAbs and JR4 mAbs) to validate the design and crystal structure of a stabilized inner domain (ID) of HIV-1 gp120. This ID displays a major antibody-dependent cellular cytotoxicity (ADCC) target of the A32 region. Current research supports a role for ADCC in preventing HIV-1 infection and in vaccine-induced protection that correlates with slower HIV progression or decreased virus replication.

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Topics: Molecular Biology, Protein Estimation, Protein Concentration, Protein Labeling, Buffers & Chemicals

Exposure to Intestinal microbes in early life can influence development of thymic lymphocytes : A review of current literature

Posted by The Protein Man on Jun 23, 2020 10:11:22 AM


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Topics: Western Blotting, Protein Electrophoresis, Protein Estimation, Protein Concentration, Protease Inhibitors, Protein Fractionation, Protein Labeling

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